ABSTRACT
High doses of chemotherapeutics in clinical treatment, leading to cell toxicity, can be lowered by co-administration of other immunoregulatory drugs. The aim of this study was to evaluate effects of several derivatives of cyclolinopeptide A (CLA), derived from linen seeds, on the suppressive action of metothrexate (MTX) in a mouse model of humoral immune response in vitro. New CLA analogues 1 and 2, and their linear precursors 3 and 4, containing conformationally restricted dipeptide fragment Phe-Phe or D-Phe-D-Phe with ethylene bridge (-CH(2)-CH(2)-) between phenylalanine nitrogens were synthesized. NMR studies and theoretical calculations showed that introduction of locally constraining fragment into CLA molecule increased its overall conformational flexibility. The bioactivity of new CLA analogues was examined in the mouse model of the in vitro secondary humoral immune response, suppressed by methotrexate (MTX). The results revealed differential actions of the peptides such as 1/augmentation of the suppressive activity of MTX or 2/antagonistic effects of the peptides on MTX-induced suppression. Potential advantages for the application of CLA-derived peptides in therapy and structure-activity relationships were discussed.
Subject(s)
Immunity, Humoral/drug effects , Methotrexate/pharmacology , Peptides, Cyclic/pharmacology , Animals , Female , Magnetic Resonance Spectroscopy , Mice , Mice, Inbred BALB C , Models, Molecular , Molecular Dynamics Simulation , Peptides, Cyclic/chemistryABSTRACT
Alpha-cyclodextrin was shown to be convenient chemical shift reagent for determination of the enantiomeric composition of alpha-hydroxyphosphonic acids by means of 31P NMR. The developed methodology appeared to be reliable, repetitive, easy to perform and simple for interpretation. Enantiomeric discrimination in the 31P NMR spectra for 12 of 13 studied hydroxyphosphonates was achieved, with baseline separation of resonances obtained for eight compounds. In those cases, the chemical nonequivalence values ranged from 0.069 to 0.313 ppm. The studies showed that enantioselectivity is strongly influenced by the solution pD and the optimal condition was found at pD 2 or 10 depending on the guest structure. On the basis of the ROESY spectra the complexation modes of selected hydroxyphosphonates with alpha-cyclodextrin was postulated.
Subject(s)
Magnetic Resonance Spectroscopy/methods , Organophosphonates/chemistry , alpha-Cyclodextrins/chemistry , StereoisomerismABSTRACT
The influence of aqueous solution pD on stereoselective complexation of N-benzyloxycarbonylaminophosphonic acids with alpha-cyclodextrin was investigated by means of nuclear magnetic resonance spectroscopy. The highest enantiodiscrimination was achieved at pD close to the pKa of less acidic hydroxyl group of the phosphonic moiety of analytes (6.5-7.5). This effect results from the stereoselective differentiation of pKa (up to 0.28 pD unit) upon complexation with applied chemical shift reagent. Moreover, analysis of 2D-ROESY spectra proved that the host-guest inclusion mode is strongly influenced by pD.
Subject(s)
Organophosphonates/chemistry , alpha-Cyclodextrins/chemistry , Hydrogen-Ion Concentration , Magnetic Resonance Spectroscopy , Molecular Structure , Solutions , Stereoisomerism , WaterABSTRACT
Capillary electrophoresis was successfully applied for separation of the enantiomers of N-benzyloxycarbonyl-alpha-aminophosphonic and alpha-aminophosphinic acids as well as their ethyl and phenyl monoesters with the use of a range of commercially available cyclodextrins (alpha, beta and hydroxypropyl-gamma-cyclodextrins) as chiral selectors. The dependence of effectiveness of separation on type and concentration of these chiral selectors as well as on pH of background electrolyte was examined in some detail.
Subject(s)
Cyclodextrins/chemistry , Electrophoresis, Capillary/methods , Organophosphonates/chemistry , Phosphinic Acids/chemistry , Esters , Hydrogen-Ion Concentration , Molecular Structure , Reproducibility of Results , StereoisomerismABSTRACT
Indirect UV detection capillary electrophoresis (CE) was used for the separation of aminoalkanephosphonic acid (AP) enantiomers by applying commercially available cyclodextrins as chiral discriminators. The results show that the separation of the enantiomers depends on pH of the background electrolyte, the molar ratio of cyclodextrin to aminophosphonic acid, and on the type of the applied chiral selector. Optimization of process conditions allowed enantiomeric baseline separation or partial separation of 12 out of 14 alpha-aminophosphonic acids studied. This type of CE might therefore be successfully used for routine determination of enantiomeric purity of aminophosphonic acids.
Subject(s)
Electrophoresis, Capillary/methods , Organophosphonates/isolation & purification , Amines/isolation & purification , Cyclodextrins , Hydrogen-Ion Concentration , StereoisomerismABSTRACT
The detailed studies concerning capillary electrophoresis separation of aminophosphonic acid enantiomers with various commercially available cyclodextrins are presented. The obtained results show that the separation of these stereoisomers is dependent on pH of background electrolyte, concentration of cyclodextrin as well as on the type of applied chiral selector. The separation mechanism is based on the co-operative effect of hydrogen bond type interactions enhanced by hydrophobic forces and sterical constrains between aminophosphonate and cyclodextrin. With application of elaborated method, enantiomeric baseline or partial separation of 18 alpha-aminophosphonic acids was achieved. This separation can be successfully used for routine aminophosphonic acids enantiopurity determination.
